The Effects of Different Methods of Anaesthesia for Laparoscopic Radical Gastrectomy with Monitoring of Entropy.

AIM: To investigate the effects of methods of total intravenous anaesthesia (TIVA) and combined intravenous and inhaled anaesthesia (CIIA) for laparoscopic radical gastrectomy under the same anaesthetic depth monitored by entropy indices. PATIENTS AND METHODS: One hundred patients undergoing laparoscopic radical gastrectomy were randomly distributed into group I (anaesthetized by TIVA) and group II (anaesthetized by CIIA), each group including 50 patients. TIVA was performed with propofol and remifentanil by means of target controlled infusion (TCI) for the patients in group I. CIIA was performed for patients in group II by inhalation of sevoflurane and continuous infusion of remifentanil after anaesthesia induction, with state entropy (SE) maintained in the range of 45-60 and difference regarding response entropy (RE) and SE less than 10.3. The concentrations of epinephrine, norepinephrine and dopamine in plasma from radial artery blood samples were measured and the durations of surgical operation, breathing recovery, extubation, awakening, and postoperative orientation recovery recorded; and 48 h postoperative adverse reactions at the following times: the time at which the patient becomes calm for 5 min after entering the operating theatre (T0); upon completion of pneumoperitoneum (PPT) (T1); 15 min after PPT (T2); intraoperative detection (T3), immediately after extubation (T4); and 15 min after extubation (T5). RESULTS: Comparing the measurements of epinephrine, norepinephrine and dopamine in plasma of the above two groups at the same time, the difference between the measurements at T0 and T2, and T5 were not statistically significant (p>0.05), whereas those at T1, T3, and T4 were statistically significant (p<0.05). Specifically, the measurements for group I were significantly higher than those for group II; the differences regarding the duration of breathing recovery, extubation, and awakening in both groups were not statistically significant (p>0.05). The postoperative orientation recovery duration for group II was significantly less than that that for group I (p<0.05); none of the patients in either group had intraoperative awareness, and the incidence of adverse reactions, such as nausea, vomit, and agitation in both groups was not statistically significantly different (p>0.05). CONCLUSION: At the same anaesthetic depth, the CIIA method outperforms the TIVA method in suppressing the stress response and obtaining smooth awakening after laparoscopic radical gastrectomy for patients with gastric cancer; therefore, the CIIA method has a better anaesthetic effect.

[Effect of comprehensive care for chronic filariasis patients with lymphedema].

OBJECTIVE: To evaluate the effect of chronic filariasis patients with lymphedema after comprehensive cared. METHODS: A total of 386 chronic filariasis patients with lymphedema received the comprehensive care including soaking feet by TCM, washing feet by single Chinese medicine or clear water, preventing and eliminating invasive wound, physical training, raising the limb, and wearing suitable shoes. The attack frequency of inflammation of lymphatic vessels, the stage of lymphedema disease, and leg circumference were observed before and after the care. RESULTS: After the comprehensive care, the attack rates of inflammation of lymphatic vessels decreased from 9.65% to 4.67%, and there was a significant difference (χ2 =7.34, P=0.0067). The proportion of the patients with high stage of lymphedema disease decreased significantly (all P <0.01), and the average leg circumference decreased of 1.06 cm. The skin appearance improved significantly. Other signs, such as pitting edema, ulcer, and skin folds also improved significantly (all P <0.01). CONCLUSION: The comprehensive care for chronic filariasis patients with lymphedema has a remarkable effect and is worthy of further application.

MicroRNA-873 Inhibits Morphine-Induced Macrophage Apoptosis by Elevating A20 Expression.

OBJECTIVES: To study the role of microRNA-873 (miR-873) in suppressing morphine-induced macrophage apoptosis and morphine dependence, and to identify molecular targets within the miR-873 pathway for the treatment of immune suppression and morphine addiction. METHODS: As morphine elevates TLR9 expression and induces TLR9-mediated apoptosis, we used TLR9 knockout Balb/C mice to study TLR9-independent effects of miR-873 on morphine-induced macrophage apoptosis. Forty TLR9-knockout mice were randomly and equally assigned to morphine group and control group. Following the administration of morphine, miR-873 mimics or miR negative control was injected into mice in each group. Using freshly isolated macrophages from mice and RAW264.7 murine macrophage cell line, miR-873 level was determined by qRT-PCR and morphine induced apoptosis was measured by TUNEL assays. Western blotting was used to detect and quantify the expression level of A20, a protein that negatively regulates inflammation and TNF-induced apoptosis. RESULTS: qRT-PCR analysis revealed a markedly lower expression of miR-873 in freshly isolated peritoneal macrophages, liver tissue and spleen tissue in the morphine group compared with the corresponding tissues in the control group. TUNEL assays showed that the apoptosis rates in the morphine groups treated with miR-873 mimics was markedly lower than their respective control groups. Western blotting results showed that A20 expression level was sharply elevated in the experimental groups treated with miR-873 mimics than the negative and blank control groups. CONCLUSIONS: Our results show that miR-873 elevates A20 levels and inhibits morphine-induced macrophage apoptosis.

[Prognostic value of PSA kinetics in locally advanced prostate cancer treated by maximal androgen blockade combined with brachytherapy].

OBJECTIVE: To evaluate the effect of post-treatment PSA kinetics on the prognosis of prostate cancer (PCa). METHODS: We retrospectively reviewed the clinical data of 114 cases of locally advanced PCa treated by maximal androgen blockade (MAB) combined with brachytherapy, and analyzed the association of the changes in PSA kinetics with the prognosis of the patients. RESULTS: The median survival time of the patients was 81 (15 - 144) months, with 1-, 3- and 5-year survival rates of 91. 23%, 78.07% and 68.42% , respectively. Univariate analysis indicated that the baseline PSA level, PSA nadir, the time of PSA decreasing to nadir, PSA doubling time, and the extent of PSA declining were all predictive factors for the survival time of the PCa patients. Multivariate analysis demonstrated that PSA nadir, the time of PSA decreasing to nadir, and the extent of PSA declining were three independent prognostic factors, which prolonged the long-term survival of the patients by 1.7, 3.2 and 6.8 times, respectively. CONCLUSION: For locally advanced PCa treated by MAB combined with brachytherapy, PSA nadir <1 micro g/L, the time to nadir <3 months, and the extent of PSA declining >96% are independent prognostic factors.

Microbial synthesized biodegradable PHBHHxPEG hybrid copolymer as an efficient intracellular delivery nanocarrier for kinase inhibitor.

BACKGROUND: Protein Kinases are key regulators of cell function and play essential roles in the occurrence and development of many human diseases. Many kinase inhibitors have been used for molecular targeted treatment of those diseases such as cancer and inflammation. However, those highly hydrophobic kinase inhibitors shared the common features of poor bioavailability and limited in vivo half-life, which strongly impeded their practical applications. Our previous study demonstrated that microbial synthesized biodegradable polyester poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx), a member of polyhydroxyalkanoates (PHAs) family, could serve as a promising delivery nanocarrier for those hydrophobic kinase inhibitors. Recently, a novel natural synthesized hybrid copolymer, PEG200 end-capped PHBHHx (PHBHHxPEG) was produced by Aeromonas hydrophila fermentation. In this study, the novel PHBHHxPEG NPs were prepared and investigated to serve as intracellular delivery nanocarriers for sustained release of hydrophobic kinase inhibitors. RESULTS: PHBHHxPEG nanoparticles (NPs) prepared by an emulsification-solvent evaporation method were spherical with a diameter around 200 nm. The entrapment efficiency on rapamycin in PHBHHxPEG NPs was 91.9% and the sustained release of rapamycin from PHBHHxPEG NPs could be achieved for almost 10 days. The cellular uptake of PHBHHxPEG NPs was significant higher than that of PHBHHx NPs. The anti-proliferation effect and mTOR inhibition ability of rapamycin-loaded PHBHHxPEG NPs was stronger than that of drug-loaded PHBHHx NPs and free rapamycin. CONCLUSIONS: PHBHHxPEG NPs could achieve the efficient entrapment and sustained release of rapamycin. The novel biodegradable PHBHHxPEG appeared a promising nanocarrier for sustained delivery of hydrophobic kinase inhibitors with improved cellular uptake and kinase inhibition efficiency.

Epithelial-mesenchymal transition and migration of prostate cancer stem cells is driven by cancer-associated fibroblasts in an HIF-1α/ß-catenin-dependent pathway.

Although cancer stem cells (CSCs) play a crucial role in seeding the initiation of tumor progression, they do not always possess the same potent ability as tumor metastasis. Thus, precisely how migrating CSCs occur, still remains unclear. In the present study, we first comparatively analyzed a series of prostate CSCs, which exhibited a dynamically increasing and disseminating ability in nude mice. We observed that the transcriptional activity of HIF-1α and ß-catenin became gradually elevated in these stem cells and their epithelial-mesenchymal transition (EMT) characteristic altered from an epithelial type to a mesenchymal type. Next, we further used cancer-associated fibroblasts (CAFs), which were cultured from surgically resected tissues of prostate cancer (PCa) to stimulate prostate CSCs. Similar results were reconfirmed and showed that the protein levels of both HIF-1α and ß-catenin were markedly improved. In addition, the EMT phenotype displayed a homogenous mesenchymal type, accompanied with increased aggressive potency in vitro. Most importantly, the aforementioned promoting effect of CAFs on prostate CSCs was completely repressed after "silencing" the activity of ß-catenin by transfection of stem cells with ShRNA. Taken together, our observations suggest that prostate migrating CSCs, with a mesenchymal phenotype, could be triggered by CAFs in a HIF-1α/ß-catenin-dependent signaling pathway.

[Epithelial-mesenchymal transition of prostate cancer: cancer stem cells or bulk cancer cells].

OBJECTIVE: To test the hypothesis that epithelial-mesenchymal transition (EMT) of prostate cancer is most likely to occur in cancer stem cells (CSC). METHODS: The isolation of CSC from LNCaP cell line was performed by flow cytometry based on side-population (SP) phenotype. After SP sorting, LNCaP/SP and LNCaP/NSP were used for further transfection of hypoxia-inducible factor-1α (HIF-1α). Subsequently, EMT-associated proteins were detected by Western blotting. And the assays of Transwell and methyl thiazolyl tetrazolium (MTT) were used to compare invasive and proliferative potency between LNCaP/SP and LNCaP/NSP after HIF-1α induction. Eventually, xenograft experiments were performed with LNCaP/HIF-1α/SP and LNCaP/HIF-1α/NSP cells for further analysis of in vivo tumorigenesis and distant metastasis. RESULTS: Through HIF-1α-induced EMT, LNCaP/HIF-1α/SP exhibited such remarkable EMT characteristics as a positive expression of epithelial markers (E-cadherin and CK18) and a negative expression of mesenchymal markers (vimentin, N-cadherin, fibronectin, cathepsin D, MMP-2 and uPAR). And LNCaP/HIF-1α/NSP underwent partial EMT with an abnormal expression of some mesenchymal proteins (vimentin and cathepsin D) and loss of epithelial protein (CK18) despite reservation of another important epithelial marker (E-cadherin). Further Transwell and MTT assays indicated that LNCaP/HIF-1α/SP exhibited stronger in vitro invasive and proliferative potency than LNCaP/HIF-1α/NSP cells. In animal models, the volume of subcutaneous tumor by LNCaP/HIF-1α/SP cells was much greater than that by LNCaP/HIF-1α/NSP counterparts ((1008 ± 230) vs (288 ± 145) mm(3), P < 0.01). Moreover, LNCaP/HIF-1α/SP cells also had a significantly higher rate of subcutaneous tumor incidence (80% vs 53%, P < 0.05) and bone metastasis (40% vs 0, P < 0.01) as compared with LNCaP/HIF-1α/NSP counterparts. CONCLUSION: As the main target cells of prostatic EMT, CSCs may develop a more malignant phenotype after EMT.

[Long-term outcomes of prostate cancer patients treated with combined brachytherapy and maximal androgen blockade].

OBJECTIVE: To explore the prognostic factors of prostate cancer (PCa) patients and evaluate the effect of brachytherapy on survival time. METHODS: A total of 289 PCa were recruited to collect their clinical and survival data. And their possible prognostic factors were analyzed. A further comparison of 5-year cumulative survival rate was made between the patients treated by maximal androgen blockade (MAB) and those on MAB plus brachytherapy. RESULTS: Their median survival time was 73 (7-144) months. And the 1, 3 and 5-year survival rates were 93.1%, 81.0% and 60.2% respectively. Univariate analysis indicated that prostate volume, basal level of prostate-specific antigen (PSA), Gleason score, tumor stage, PSA nadir, time PSA decreasing to nadir and brachytherapy were all predictive factors for survival time. And multivariate analysis further demonstrated that Gleason score, tumor stage and PSA nadir were independent prognostic indicators. And the combination therapy based on brachytherapy could significantly increase the 5-year cumulative survival rate than MAB-based monotherapy. CONCLUSION: Gleason score, tumor stage and PSA nadir may predict the prognosis of PCa patients. And brachytherapy significantly improves patient survival.

[Longitudinal surveillance of intestinal nematodiasis in Yinghu Village of Wujiang City].

OBJECTIVE: To understand the epidemic dynamics of intestinal nematodiasis in Wujiang City. METHOD: The residents of Yinghu Village of Wujiang City were investigated with Kato-Katz technique for the infections of intestinal nematodes, and the persons with the infections were administered with anthelmintics and they also received the health education. RESULTS: Of the 5 757 residents, 147 persons were infected with intestinal nematodes, with the average infection rate of 2.55%. The infection rates of hookworm, Ascaris lumbricoides and Tricuris trichiura were 1.96%, 0.49% and 0.24%, respectively. The repeated infection rate was 20.30%, that was 10.36 times higher than the new infection rate (1.96%). CONCLUSIONS: There are still regions and populations with high infection rates of intestinal nematodes in Wujiang City where the intestinal nematodiasis has been controlled. Therefore, it is necessary to strengthen the monitoring, prevention and control work.

Heterologous expression of human costimulatory molecule B7-2 and construction of B7-2 immobilized polyhydroxyalkanoate nanoparticles for use as an immune activation agent.

BACKGROUND: Costimulation of T cells via costimulatory molecules such as B7 is important for eliciting cell-mediated antitumor immunity. Presenting costimulation molecules by immobilizing recombinant B7 on the surface of nanovectors is a novel strategy for complementary therapy. Polyhydroxyalkanoates (PHAs) are a family of biodegradable, non-toxic, biocompatible polyesters, which can be used as a nonspecific immobilizing matrix for protein presentation. Recombinant protein fusion with PHA granule binding protein phasin (PhaP) can be easily immobilized on the surface of PHA nanoparticles through hydrophobic interactions between PhaP and PHA, and therefore provides a low-cost protein presenting strategy. RESULTS: In this study, the extracellular domain of the B7-2 molecule (also named as CD86) was fused with PhaP at its N-terminal and heterogeneously expressed in recombinant Escherichia coli strain BL21 (DE3). The purified B7-2-PhaP protein was immobilized on the surface of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx)-based nanoparticles. Loading of 240 µg (3.2 pMol) of B7-2-PhaP protein per mg nanoparticles was achieved. Immobilized B7-2-PhaP on PHBHHx nanoparticles induced T cell activation and proliferation in vitro. CONCLUSIONS: A PHA nanoparticle-based B7-2 costimulation molecule-presenting system was constructed. The PHA-based B7 presenting nanosystem provided costimulation signals to induce T cell activation and expansion in vitro. The B7-2-PhaP immobilized PHA nanosystem is a novel strategy for costimulation molecule presentation and may be used for costimulatory molecule complementary therapy.

Social determinants of health and depression: a preliminary investigation from rural China.

BACKGROUND: In the last several years, research related to social determinants of health (SDH) has begun to resonate in the medical, behavioral, social and political sciences arena. The aim of the present study was to explore the relationship between SDH and depression, and to provide new evidences and clues for depression control and prevention. METHODOLOGY/PRINCIPAL FINDINGS: This research was a cross-sectional survey executed door to door from October 2006 to April 2008, with a sample of 3,738 individuals aged 18 and older in rural China. The three variables of SDH were socioeconomic status (years of schooling and self-reported economic status of family), social cohesion and negative life events. Demographic variables and self-perceived physical health were taken as potential confounders. The cross-table analysis showed that variations in levels of depression were associated with variations in SDH, and logistic regression analysis confirmed the association even after adjusting for potential confounding variables. CONCLUSIONS: Although there were some limitations, the current study provides initial evidence of the importance of SDH in depression. Findings indicate that social inequity and the role of policy action emphasized by SDH should be considered high priorities when addressing the issue of depression. In addition, cell-to-society and pill-to-policy approaches should be encouraged in the future.

[Sorting and identification of cancer stem cells in human prostate cancer cell lines].

OBJECTIVE: To sort and identify side population (SP) cancer stem cells (CSC) in human prostate cancer (PCa) cell lines. METHODS: Stem-like cells were isolated from five PCa cell lines Du145, IA8, LNCaP, TSU-Pr and PC-3 using FACS based on CD133+ CD44+ immunophenotype and SP in Hoechst staining. The in vitro growth pattern and tumorigenicity of SP stem cells were verified by soft agar colony-formation trial. LNCaP/SP cells were selected for further identification of stem cell properties using immunostaining, proliferation and invasion assay. Eventually, tumorigenicity and metastasis ability of LNCaP/SP were confirmed by xenograft experiments. RESULTS: The percentages of CSCs of the CD133 CD44 + immunophenotype were extremely low in the five PCa cell lines. On the contrary, the percentages of the isolated SP cells were significantly higher in Du145 ([0.15 +/- 0.02]%), IA8 ([0.60 +/- 0.07 ]%), LNCaP ([0.8 +/- 0.1]%) and TSU-PrL ([2.0 +/- 0.4]%), but none was detected in PC-3. Besides, IA8/SP, LNCaP/SP and TSU-PrL/SP cells showed a significantly greater colony-forming efficiency than non-side population (NSP) cells (P < 0.05). Compared with LNCaP/NSP cells, LNCaP/SP cells exhibited high expressions of integrin alpha2, Nanog, CD44, OCT4 and ABCG2, remarkably enhanced invasive and proliferative potentials in vitro, and markedly increased tumorigenicity and metastasis (P < 0.01). CONCLUSION: SP sorting is more suitable than CD133+ CD44+ selection for enriching CSCs from PCa cell lines, and LNCaP/ SP represents a typical CSC population.

Effects of prostate manipulation on serum total and free prostate specific antigen, and free-to-total prostate specific antigen ratio.

OBJECTIVE: To evaluate the effects of the different types of manipulation on prostate total specific antigen (tPSA), free prostate specific antigen (fPSA), and free-to-total prostate specific antigen (f/tPSA). METHODS: A total of 160 males were enrolled from January 2006 to December 2009 in the Urology Department, Beijing Anzhen Hospital affiliated to the Capital Medical University, Beijing, China. Of these patients, 23 had digital rectal examination (DRE), 21 had urethral catheterization, 28 had rigid cystoscopy, 35 had prostate biopsy, 35 underwent transurethral resection of the prostate (TURP), and 18 underwent suprapubic prostatectomy. Blood samples were taken before, at 24 hours, and 4 weeks after the manipulation for PSA tests. RESULTS: The DRE had no significant effect on PSA. Catheterization and cystoscopy exerted significant increases in tPSA at 24 hours. However, these small increases may not be clinically significant. The fPSA and f/tPSA were not significantly changed. There was a marked increase in tPSA and fPSA, associated with a decrease in f/tPSA at 24 hours after biopsy. No significant alterations were found in tPSA, fPSA, and f/tPSA at 4 weeks after catheterization, cystoscopy, and biopsy. The TURP and prostatectomy caused significant increases in tPSA and fPSA at 24 hours, associated with decreases in f/tPSA. The tPSA and fPSA values were below the baseline levels at 4 weeks after TURP and prostatectomy, however, f/tPSA remained constant. CONCLUSION: The DRE, catheterization, and cystoscopy had no crucial effect on PSA. Prostatic biopsy, TURP and prostatectomy significantly affected the PSA levels, and their longitudinal courses should be considered while evaluating different forms of PSA levels.

[Finasteride: an effective therapeutic for benign prostatic hyperplasia related gross hematuria in patients receiving anticoagulant].

OBJECTIVE: To study the effect of finasteride on benign prostatic hyperplasia (BPH) related gross hematuria in patients receiving anticoagulant. METHODS: A total of 105 patients with BPH related gross hematuria were divided into an anticoagulant group (n = 81), treated with combined therapy of anticoagulants and finasteride, and a control group (n = 24), given finasteride only at 5 mg daily. The therapeutic effects were compared by a 6-month follow-up. RESULTS: In the anticoagulant group, gross hematuria was cured in 52 patients (64.2%), taking an average time of 3.9 weeks (1-6 weeks), and improved in 12 patients (14.8%), as compared with 16 patients cured (66.7%), 3.2 weeks taken (1-5 weeks), and 4 patients improved (16.7%) in the control group. The mean time taken to resolve hematuria was longer in the former (P < 0.05). But the cure rates had no significant differences either between the two groups or among the subgroups receiving different anticoagulants. CONCLUSION: Finasteride is an effective therapeutic for BPH related hematuria in patients receiving different anticoagulants. It makes no significant differences in cure and effectiveness rates between patients treated with and without anticoagulant, but takes an average of longer time to resolve hematuria in patients receiving anticoagulant.

[Over-expression of HIF-1alpha induces EMT of human prostate cancer cells].

OBJECTIVE: To determine whether human prostate cancer cell lines undergo epithelial-mesenchymal transition (EMT) and become more invasive when induced by HIF-1alpha, and to explore the underlying molecular mechanism. METHODS: The cell line LNCaP, appropriate for the HIF-1alpha induction test, was screened out from 4 different EMT-negative prostate cell lines through vimentin gene detection by RT-PCR. The recombinant plasmid pCDNA3. 1(-)/HIF-1alpha was constructed and transfected into LNCaP with the Lipofectamine 2000 system. The control plasmid pCDNA3.1 (-) was transfected by the same method. The positive clone cells were selected by G418 and confirmed by Western blot and immunofluorescence staining. Then a Transwell polycarbonate filter, coated with 100 micol Matrigel at 1:20 dilution in the serum-free medium, was used to analyze the invasive potency. The expression of E-cadherin and vimentin was detected by Western blot. RESULTS: Among the 4 different EMT-negative cell lines, LNCaP was the only one that expressed the vimentin gene but not protein. The expression of HIF1alpha was obviously higher in LNCaP/HIF1alpha than in LNCaP/pCDNA3. 1 (- an LNCaP. The number of the LNCaP/HIF1alpha cells that penetrated through the Transwell polycarbonate filter was significantly larger than that of the LNCaP and LNCaP/pCDNA3. 1(-) cells. Compared with the LNCaP/pCDNA3.1(-) and LNCaP cells, the expression of vimentin was up-regulated, while that of E-cadherin down-regulated, in LNCaP/HIF1alpha. CONCLUSION: The over-expression of HIF-1alpha could induce EMT in the human prostate carcinoma cell line LNCaP and enhance its invasiveness through E-cadherin and vimentin regulation.

Resveratrol reestablishes spermatogenesis after testicular injury in rats caused by 2, 5-hexanedione.

BACKGROUND: Environmental toxins can destroy the physiological process of spermatogenesis and even lead to male infertility. Resveratrol (RES) is a natural phytoalexin with a wide range of biological activities. Some recent researches have demonstrated that RES can increase sperm output and protect sperm from apoptosis caused by physical damage. However, there is no evidence indicating that it can also exhibit a similar activity in testis injury caused by environmental toxins. This study was designed to evaluate the protective effect of resveratrol on testis damaged by environmental toxins and to elucidate the possible mechanism of its protective effect. METHODS: In this study 2, 5-hexanedione (2, 5-HD) was used as the injury agent. Forty male SD rats were randomly divided into 5 groups. During the first 5 weeks, group A was raised normally, groups B, C, D and E were exposed to 1% 2, 5-HD; during the following 9 weeks, group C, D, E received intragastric administration of different concentrations of resveratrol (20 mg x kg(-1) x d(-1), 40 mg x kg(-1) x d(-1) and 80 mg x kg(-1) x d(-1)), while groups A and B were treated by carboxymethylcellulose. Physical signs, body weight gain and testis weight were comparatively observed. Numbers and diameters of seminiferous tubules were analyzed following HE staining. In addition, expression of the c-kit protein and gene in spermatogenic cells in every group was detected with immunohistochemistry, Western blot or RT-PCR. RESULTS: The 2, 5-HD treatment resulted in physical signs that became worse and in emarciated testis. HE staining and immunohistochemistry showed that seminiferous tubules became emarcid, obsolete spermatogonia being stagnant and expression of c-kit protein being depressed. After oral administration of resveratrol, the 2, 5-HD-induced physical signs were improved and close to the normal rats. The gain of body weight increased (P < 0.01). The recovery of testis weight was significant (P < 0.01). At the histological level, the seminiferous epithelia began to differentiate (P < 0.01); and even the physiological process of spermatogenesis restarted. Moreover, expression of c-kit protein and gene function resumed, although its expression remained different from the normal group. The diameter of and number of seminiferous tubules and the expression level of c-kit protein and gene activity were much closer to the normal group with increased doses of the resveratrol through oral administration. CONCLUSIONS: Resveratrol could ameliorate markedly the dyszoospermia induced by 2, 5-HD and induce spermatogenesis. The expression of c-kit, which is a specific marker protein of spermatogenic cell membranes, could be regulated by resveratrol.

[Loss of heterozygosity on chromosome 10 is associated with bone metastasis of human prostate cancer].

OBJECTIVE: To study the cytogenetic mechanism of bone metastasis of human prostate cancer (PCa). METHODS: We analyzed chromosome variation by comparative genomic hybridization in 18 patients with prostate cancer to determine the chromosome variants associated with bone metastasis, and focused on 7 microsatellite sites on chromosome 10 for the detection of the loss of heterozygosity (LOH) by PCR-based microsatellite polymorphism analysis. RESULTS: In the 11 samples with bone metastasis, the variation rate of chromosome 10 was 90.9% (10/11), significantly higher than that of the others (P < 0.01). A much higher LOH frequency was observed at the 7 microsatellite loci on chromosome 10 and the highest located in 10q24. 2-q25.3 (D10S1693-D10S587) in the PCa patients with bone metastasis. CONCLUSION: There is a high-frequency LOH region in 10q24. 2-q25.3 (D10S1693-D10S587) on chromosome 10 in PCa patients with bone metastasis, which may be potentially involved in PCa progression and specific bone metastasis.

[Expression of coxsackie B virus-adenovirus receptor in prostate cancer Du145 and LNCaP cell lines and its significance].

OBJECTIVE: To observe the expression of Coxsackie B virus-adenovirus receptor (CAR) in two prostate cancer cell lines with different metastatic potentials. METHODS: The expressions of CAR in two prostate cancer cell lines (Du145 and LNCaP) with different metastatic potentials were detected by Western blotting. The Transwell polycarbonate filter was used to analyze the invasive potency. RESULTS: As one of the adhesion associated proteins, CAR highly expressed in the LNCaP cell line, which is well known with a low metastatic potential, and lowly expressed in Du145 with a high metastatic potential (P < 0.01). The invasive potency of Du145 was significantly higher than that of LNCaP (P < 0.05). CONCLUSION: There was a difference in the metastatic phenotypes of CAR among cell lines with different metastatic potentials. The expressions of CAR proteins may play an important role in repressing the metastasis of prostate cancer.

Role of Wnt/beta-catenin signaling pathway in epithelial-mesenchymal transition of human prostate cancer induced by hypoxia-inducible factor-1alpha.

OBJECTIVES: Epithelial-mesenchymal transition (EMT) is an important process in tumor development, and several studies suggest that the Wnt/beta-catenin signal pathway may play an important role in EMT. However, there is no direct evidence showing that the Wnt/beta-catenin pathway actually determines the EMT induced by an exogenous signal. Our previous research has successfully proved that overexpression of hypoxia-inducible factor-1alpha (HIF-1alpha) could induce EMT in LNCaP cells, but not in PC-3. The present study aims to determine whether the signal of HIF-1alpha for inducing prostate cancer cells to undergo EMT might possibly pass through the Wnt/beta-catenin pathway. METHODS: Epithelial-mesenchymal transition associated proteins were detected in several human prostate carcinoma cell lines by Western blot, and then we distinguished the EMT positive cell lines from the EMT negative cell lines. Furthermore, we evaluated the possible correlation between potency of invasiveness and proliferation among these cell lines with different characteristics of EMT using Matrigel transwell and thiazolyl blue tetrazolium bromide (MTT) assays. Finally, the different expression of some critical proteins and genes in Wnt/beta-catenin signaling pathway were analyzed by Western blot and reverse transcription-polymerase chain reaction (RT-PCR) in these cells with different characteristics of EMT. RESULTS: Among several prostate cancer cell lines, PC-3, LNCaP and PC-3/HIF-1alpha are EMT negative cell lines, whereas LNCaP/HIF-1alpha and IA8 have undergone the EMT process. EMT positive cells (LNCaP/HIF-1alpha and IA8) exhibit much stronger potency of invasiveness and proliferation than those of PC-3 and LNCaP, which belong to EMT negative cells. Interestingly, although PC-3/HIF-1alpha had not completed the EMT process, it still displayed stronger potency of invasion and proliferation, resembling EMT positive cells. The protein expression level of total glycogensynthase kinase 3beta (GSK-3beta) and phospho-GSK-3beta in LNCaP/HIF-1alpha, IA8 and PC-3/HIF-1alpha cells significantly decreased; however, the relative ratios of p-GSK3beta/t-GSK3beta in LNCaP/HIF-1alpha, IA8 and PC-3/HIF-1alpha cells were significantly higher than PC-3 and LNCaP. Consistently, beta-catenin protein expression increased in LNCaP/HIF-1alpha and IA8 cells, but not in PC-3/HIF-1alpha; RT-PCR confirmed these results, except for the enhanced transcription activity of beta-catenin mRNA in PC-3/HIF-1alpha. CONCLUSION: Our data suggests that activation of the Wnt/beta-catenin signaling pathway correlates with the characteristic of EMT and potency of invasiveness and proliferation. This may be the critical factor that directly controls the process of EMT induced by HIF-1alpha in prostate cancer cells.

[Resveratrol helps restore spermatogenesis after testis injury induced by 2,5-hexanedione].

OBJECTIVE: To study the effect of resveratrol on spermatogenesis after 2,5-hexanedione(2,5-HD)-induced testicular injury. METHODS: Forty male SD rats were randomly divided into 5 groups. Group A were normally raised and Group B, C, D and E exposed to 1% 2,5-HD for 5 weeks, followed by administration of resveratrol of different concentrations (20, 40 and 80 mg/[ kg x d], respectively) to Group C, D and E for 9 weeks. Then the rats were killed, their physical signs, body weight gain and testis weight were assessed, and immunohistochemistry and Western blot analysis used to investigate the numbers and diameters of seminiferous tubules and the expression of c-kit protein of spermatogenic cell membrane. RESULTS: The rats exposed to 2,5-HD showed weak body, lax skin, dim color pattern, tardy body weight gain, and emaciated testis. Immunohistochemistry revealed emaciated seminiferous tubules, stagnant obsolete spermatogonia and negative expression of c-kit protein. After resveratrol administration, the 2,5-HD-induced physical signs were improved and close to normal. Compared with those of the 2,5-HD injured group, the body weight and testis weight of the resveratrol treated group increased obviously (P < 0.01); and the aliquots of the seminiferous epithelia began to differentiate and the spermatogenesis and expression of c-kit protein partly resumed (P < 0.01). With increasing dose of resveratrol, the diameters and numbers of seminiferous tubules (P < 0.01) and the expression levels of c-kit protein (P < 0.01) were gradually and significantly restored almost to normal. CONCLUSION: Resveratrol could promote the recovery of spermatogenesis after 2,5-HD-induced testicular injury.